cattletype MAP Ab is a sensitive and specific indirect ELISA for detecting antibodies to Myobacterium avium subsp. paratuberculosis (MAP) in serum, plasma, and milk samples from cattle and goats and in serum and plasma samples from sheep.

cattletype MAP Ab is for veterinary use only.

In Germany: Registered in accordance with §17c of the German Law on Animal Diseases. Registration No.: FLI-B 471

Specificity of cattletype MAP Ab in milk.|Specificity of cattletype MAP Ab in serum.|

A MAP-free negative population of 542 milk samples was tested using cattletype MAP Ab. An S/P (ratio sample/positive control) below 0.6 indicates the sample is negative for antibodies to MAP. Four samples gave false positive results, providing a diagnostic specificity for this sample set of 99.3%.|A MAP-free negative population of 147 serum samples was tested using cattletype MAP Ab. An S/P (ratio sample/positive control) below 0.4 indicates the sample is negative for antibodies to MAP. One sample gave a false positive result, providing a diagnostic specificity for this sample set of 99.3%.|

cattletype MAP Ab sensitivity was determined using 102 serum samples and 55 milk samples (samples from a variety of sources including German Reference Laboratory for Paratuberculosis, GD Animal Health Service Deventer, and State Veterinary Diagnostic Laboratory of Saxony). Of 102 positive serum samples, 92 samples tested positive for MAP-specific antibodies, providing a diagnostic sensitivity in serum samples for this sample set of 90.2% (see the table Sensitivity of cattletype MAP Ab in serum). Of 55 positive milk samples, 49 samples tested positive for MAP-specific antibodies, providing a diagnostic sensitivity in milk samples for this sample set of 89.1% (see the table Sensitivity of cattletype MAP Ab in milk).

Sensitivity of cattletype MAP Ab in serum

Samples

No. of positive samples

No. of samples scored positive with cattletype MAP Ab

Correctly scored %

A

3

3

100

B

2

2

100

C

36

34

94.4

D

9

8

88.9

E

34

34

100

F

10

6

60

G

8

5

62.5

Total

102

92

90.2

Sensitivity of cattletype MAP Ab in milk

Samples

No. of positive samples

No. of samples scored positive with cattletype MAP Ab

Correctly scored %

A

10

5

50

B

8

7

87.5

C

3

3

100

D

31

31

100

E

3

3

100

Total

55

49

89.1

Principle

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent for paratuberculosis, which is also called Johne's Disease. Paratuberculosis is an incurable and chronic infectious disease with a long incubation time, characterized by excessive weight loss and persistent diarrhea in cattle in the final stage of the disease. MAP is spread worldwide among ruminants.

cattletype MAP Ab ELISA permits the semi-quantitative detection of anti-MAP antibodies and can be used with serum, plasma, and milk samples. This product is an ELISA (enzyme-linked immunosorbent assay) with ready-to-use colored reagents. Samples are pre-incubated with a dilution buffer containing inactivated Mycobacterium phlei extract in order to minimize cross-reactions with atypical mycobacteria. The test plate is coated with inactivated MAP antigen. After transfer of the pre-incubated samples to the antigen-coated plate, MAP-specific antibodies bind to the immobilized antigen. The anti-Ruminant-IgG-HRP conjugate detects antibodies bound to the antigen. A color reaction is started by adding the substrate solution and stopped after 10 minutes. The optical density (OD) is measured in a spectrophotometer. The OD values correlate with the concentration of anti-MAP antibodies in the sample.

Procedure

Serum, plasma, and milk samples can be tested after dilution. Milk samples can be tested after skimming.

There are two protocols for this product. They differ in the length of the incubation period. Serum and plasma samples are incubated for 30 minutes with the test plate. For milk samples, an overnight incubation is needed. If antibodies to MAP are present, they will bind to the antigen on the plates. Unbound material is removed by rinsing.

The anti-IgG-HRP conjugate is added. Unbound conjugate is rinsed out.

The color reaction is started by adding the TMB Substrate Solution and stopped after 10 minutes. The optical density (OD) is measured using a spectrophotometer. The results are easy to interpret.

Applications

cattletype MAP Ab is for the detection of antibodies to Mycobacterium avium subsp. paratuberculosis in serum, plasma, and milk samples from cattle, sheep, and goats.